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通过基质辅助激光解吸电离飞行时间质谱直接鉴定需氧菌准确且可靠。

Direct Identification of Aerobic Bacteria by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Is Accurate and Robust.

作者信息

Mestas Javier, Quias Teephany, Dien Bard Jennifer

机构信息

Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California.

Keck School of Medicine at the University of Southern California, Los Angeles, California.

出版信息

J Clin Lab Anal. 2016 Sep;30(5):543-51. doi: 10.1002/jcla.21900. Epub 2015 Dec 14.

Abstract

BACKGROUND

Bacterial identification in the clinical laboratory can be laborious and expensive. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a rapid and cost-effective diagnostic tool for the identification of organisms routinely found in the microbiology laboratory. The objective of this study was to demonstrate that identification of aerobic Gram-positive and Gram-negative organisms could be performed accurately and efficiently by MALDI-TOF MS and the Bruker Biotyper system without the use of time-consuming extraction methodologies.

METHODS

Isolates previously recovered by routine culture and workup from clinical specimens were cultured to appropriate media, identified directly by MALDI-TOF MS, and compared to results from various biochemical identification methods.

RESULTS

Using the direct-smear method, 99.5% and 98.0% of aerobic Gram-negative and Gram-positive bacteria, respectively, were identified to the genus level. At a score of ≥1.9, 97.6% Gram-negative organisms and 94.6% Gram-positive organisms were correctly identified to the species level by direct-smear method. Only 1.1% of isolates required further reflex to direct-plate extraction. The direct-smear method proved to be robust, as various growth temperatures, media, culture age, and different operators had no notable impact on the bacterial identification rate.

CONCLUSION

The direct-smear method is an accurate and time-saving method for routine species-level bacterial identification.

摘要

背景

临床实验室中的细菌鉴定可能既费力又昂贵。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)是一种快速且经济高效的诊断工具,可用于鉴定微生物实验室中常见的生物体。本研究的目的是证明,使用MALDI-TOF MS和布鲁克微生物鉴定系统,无需耗时的提取方法,即可准确、高效地鉴定需氧革兰氏阳性和革兰氏阴性生物体。

方法

将先前通过常规培养和临床标本检查回收的分离株接种到合适的培养基中,通过MALDI-TOF MS直接鉴定,并与各种生化鉴定方法的结果进行比较。

结果

采用直接涂片法,分别有99.5%的需氧革兰氏阴性菌和98.0%的需氧革兰氏阳性菌被鉴定到属水平。在得分≥1.9时,通过直接涂片法,97.6%的革兰氏阴性菌和94.6%的革兰氏阳性菌被正确鉴定到种水平。仅1.1%的分离株需要进一步采用直接平板提取法。直接涂片法被证明是可靠的,因为不同的生长温度、培养基、培养时间和不同的操作人员对细菌鉴定率均无显著影响。

结论

直接涂片法是一种准确且省时的常规细菌种水平鉴定方法。

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